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Estimation of citrus canker lesion size using hyperspectral reflectance imaging

International Journal of Agricultural and Biological Engineering 2013年第3期6卷 41-51页

作者： Nikhil P.Niphadkar Thomas F.Burks Jianwei Qin Mark A.Ritenour ATW Automation Inc. DaytonOH 45402USA Department of Agricultural and Biological Engineering University of FloridaGainesvilleFL 32611USA U.S.Department of Agriculture BeltsvilleMD 20705USA Department of Horticultural Sciences University of FloridaGainesvilleFL 34945USA

The citrus industry has need for effective approaches to remove fruit with canker before they are shipped to selective international market such as the European *** research aims to determine the detectable size limit for cankerous lesions using hyperspectral imaging *** developed multispectral algorithms using visible to near-infrared wavelengths,were used to segregate cankerous citrus fruits from other peel conditions(normal,greasy spot,insect damage,melanose,scab and wind scar).However,this previous work did not consider lesion size.A two-band ratio method with a simple threshold based classifier(ratio of reflectance at wavelengths 834 nm and 729 nm),which gave maximum overall classification accuracy of 95.7%,was selected for lesion size estimation in this *** smallest size of cankerous lesion detected in terms of equivalent diameter was 1.66 *** effect of variation of threshold values and number of erosion cycles(applying morphological erosion multiple times to the image)on estimation of smallest detectable lesion was *** was found that small threshold values gave better canker classification accuracies,while exhibiting a lower overall classification ***,higher threshold values portrayed the opposite *** threshold value of 1.275 gave the optimum tradeoff between canker classification accuracy,overall classification accuracy and minimal lesion size *** the number of erosion cycles reduced detection rates of smaller canker lesions,leading to the conclusion that a single erosion cycle gave the best size estimation *** erosion kernel of the size 3 mm×3 mm was used during the exploration.

关键词： citrus canker lesion size disease detection hyperspectral reflectance imaging image classification multispectral algorithm size detection limit

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Control Effects of 9 Fungicides on citrus canker

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Agricultural Science & Technology 2021年第1期22卷 37-41页

作者： HUANG Ze-pei ZHOU Ya-lin LONG Jian-guo MO Jun-ping SUN Wei-si Plant Protection and Inspection Station of Agricultural and Rural Bureau of Mayang County Mayang 419400PRC Agricultural Technology Extension Station of Agricultural and Rural Bureau of Mayang Miao Mayang 419400PRC

In order to screen out effective fungicides for controlling citrus canker,the control effects of 9 fungicides on Jinhong bingtang orange canker were *** results showed that 77%cupric calcium sulfate WP,30%copper oxychloride SC,46%copper hydroxide WG,30%thiodiazole-copper SC and 40%zinc thiazole SC had better comprehensive control effect on citrus canker,6%benziothiazolinone WG had an average effect,3%zhongshengmycin AS,2%kasugamycin AS and 33.5%oxine-copper SC had poor control ***,fungicides could be used alternately or choose compounded *** addition,adjuvants could be considered in use,which was an effective way to enhance efficacy,reduced dosage and delayed pesticide resistance.

关键词： citrus canker Copper preparation Biological agents Thiazoles Heterocyclic Control effect Efficacy test

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Production of Transgenic Anliucheng Sweet Orange(citrus sinensis Osbeck) with Xa21 Gene for Potential canker Resistance

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Journal of Integrative Agriculture 2014年第11期13卷 2370-2377页

作者： LI Ding-li XIAO Xuan GUO Wen-wu Key Laboratory of Horticultural Plant Biology Ministry of Education/College of Horticulture & Forestry Sciences Huazhong Agricultural University College of Life and Environmental Sciences Gannan Normal University

citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR （pattern recognition receptor） gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene （HPT） was introduced into Anliucheng sweet orange （citrus sinensis Osbeck） via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin （HYG） and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identiifed by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was brielfy discussed.

关键词： citrus canker Xa21 sweet orange pattern recognition receptors (PRRs)

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Occurrence Regularity and Integrated Control Technology of canker in citrus

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Asian Agricultural Research 2021年第2期13卷 33-34,40页

作者： Zhijing WANG Liming WU Fang SONG Ligang HE Ce WANG Yingchun JIANG Institute of Fruit Tree and Tea Hubei Academy of Agricultural Sciences Wuhan 430064 China

canker is a quarantine bacterial disease that seriously harms leaves,branches and fruits of citrus,leading to a decrease in the production and affecting the commodity and sales of *** canker has the characteristics of fast spreading speed and difficult radical *** the identification of symptoms and a summary of occurrence regularity,the integrated prevention and control technology for citrus canker is described in this article,in order to achieve effective prevention and control and reduce prevention and control cost.

关键词： citrus canker Symptom recognition Occurrence regularity Integrated prevention and control

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Preparation of Monoclonal Antibody Against PthA-NLS and Cloning of the Relative ScFv Gene

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Agricultural Sciences in China 2010年第1期9卷 85-92页

作者： LI Na HU Chun-hua LONG Gui-you DAI Su-ming MA Xian-feng XIE Yu-ming DENG Zi-niu Horticulture and Landscape College Hunan Agricultural University Changsha 410128 P.R.China National Center for citrus lmprovement/Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization/Hunan Agricultural University Changsha 410128 P.R.China

The present study aimed at the preparation of monoclonal antibody against the recombinant PthA-NLS and the isolation of the relative ScFv （single chain variable fragment） genes, providing the possibility to better understand the pathogenesis mechanism via PthA, and developing proper construct for future experimentation to obtain citrus plants resistant to canker disease by transformation and plant antibody techniques. The recombinant polypeptide PthA-NLS was injected into Balb/c mice to produce monoclonal antibody. Total RNA was isolated from the hybridoma cell line 3D10H2 which secreted anti- PthA-NLS McAb, and the variable region genes were amplified with specific primers by RT-PCR and SOE-PCR （splicing by overlap extension）, and then the ScFv gene was isolated. The recombinant ScFv gene was cloned into pGEM-T and pET32a（＋） vector. The later plasmid was transferred into E. coli BL21 （DE3） and the expression of the recombinant protein was induced. Three cell lines producing monoclonal antibody against PthA-NLS were acquired and named 1C8H1, 2D12B6, and 3D8A10. The recombinant ScFv gene of about 750 bp was constructed. The sequencing results showed that the ScFv gene consists of a 360 bp heavy chain, a 342 bp light chain, and a 45 bp linker region. The recombinant fusion ScFv protein was expressed by IPTG induction, and a 44.5 kDa of recombinant fusion protein was obtained. In conclusion, we obtained three cell lines stably producing monoclonal antibody specifically bound to PthA-NLS, and the relative ScFv gene was constructed and successfully expressed in E. coli. These results may play an important role in further understanding the pathogenesis mechanism and in the development of possible citrus resistant to canker disease by genetic transformation and plant antibiobody.

关键词： citrus canker monoclonal antibody ScFv gene pthA

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The small and large subunits of carbamoyl-phosphate synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri ***

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Journal of Integrative Agriculture 2015年第7期14卷 1338-1347页

作者： GUO Jing SONG Xue ZOU Li-fang ZOU Hua-song CHEN Gong-you School of Agriculture and Biology Shanghai Jiao Tong University

Carbamoyl-phosphate synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citrisubsp. citri, carA and carB encode the small and large subunits of carbamoyl-phosphate synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hyper- sensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type Ill secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracellular enzyme activities. These data demonstrated that only the large subunit of carbamoyl-phosphate synthase was essential for canker development by X. citri subsp, citri.

关键词： Xanthomonas citri subsp citri carbamoyl-phosphate synthase pathogenicity citrus canker hrp gene

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A Potential Disease Susceptibility Gene CsLOB of citrus Is Targeted by a Major Virulence Effector PthA of Xanthomonas citri subsp, citri

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Molecular Plant 2014年第5期7卷 912-915页

作者： Zheng Li Lifang Zou Gang Ye Li Xiong Zhiyuan Ji Muhammad Zakria Ni Hong Guoping Wang Gongyou Chen School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 200240 China State Key Laboratory of Microbial Metabolism School of Life Science &Biotechnology Shanghai Jiao Tong University Shanghai 200240 China National Key Laboratory of Agromicrobiology Huazhong Agricultural University Wuhan Hubei 430070 China

Dear Editor, citrus bacterial canker （CBC）, one of the most destructive diseases of citrus, has a severe economic impact on citrus industries in tropical and subtropical countries （Das, 2003）. It has been widely known that Xanthomonas citri subsp, citri （Xcc） causes the CBC by transferring the type-Ill effectors （T3SEs） to the interior of the plant cell through type-Ill secretion system （T3S） to modulate transcription of host plants to trigger CBC disease （Schaad et al., 2006）. Pathotype A,

关键词： citrus canker citrus fruit industry NUCLEOTIDES PATHOGENIC microorganisms MUTAGENESIS

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Cloning and Expression Analysis of citrus Genes Cs GH3.1 and Cs GH3.6Responding to Xanthomonas axonopodis pv. citri Infection

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Horticultural Plant Journal 2016年第4期2卷 193-202页

作者： CHEN Min HE Yongrui XU Lanzhen PENG Aihong LEI Tiangang YAO Lixiao LI Qiang ZHOU Pengfei BAI Xiaojing DUAN Minjie JIANG Xueyou JIA Ruirui ZOU Xiuping CHEN Shanchun citrus Research Institute Southwest UniversityCitrus Research InstituteChinese Academy of Agricultural Sciences National citrus Engineering Research Center National Center for citrus Varieties Improvement

To study the functions of the early auxin-responsive genes Cs GH3.1 and Cs GH3.6 in citrus resistance against canker disease, we cloned Cs GH3.1and Cs GH3.6 in ‘Newhall' Navel Orange(citrus sinensis Osbeck). They are 1 797 bp and 1 887 bp and encode 598 and 629 amino acids, respectively. In vitro mature leaves from susceptible ‘Newhall' and resistant Calamondin(C. madurensis) were inoculated by a Xanthomonas axonopodis pv. citri(Xac) bacterial suspension, and expression of Cs GH3.1 and Cs GH3.6 in the two varieties were analyzed using quantitative real-time PCR(q RT-PCR). ‘Newhall' leaves were treated with different hormones for 3 days, inoculated by Xac bacterial suspension, and then the symptoms in these leaves were investigated. We used q RT-PCR to analyze the effect of different hormones on Cs GH3.1 and Cs GH3.6 expression in ‘Newhall'leaves. The expression levels of both Cs GH3.1 and Cs GH3.6 were significantly induced by Xac in ‘Newhall' leaves, compared with levels in Calamondin leaves. 1-naphthy acetic acid(NAA) increased the hypertrophy of infection sites in ‘Newhall' leaves, while naphthyl-phthalamic acid(NPA)had no visible effect on lesion development. NAA hormone greatly improved expression of Cs GH3.1 in ‘Newhall', but not Cs GH3.6. These results indicate that the auxin primary-response gene Cs GH3.1 plays an important role in citrus susceptibility to Xac.

关键词： citrus citrus canker auxin signal transduction Cs GH3 1 Cs GH3 6

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NADPH oxidase might negatively feedback regulate the generation of H2O2 in citrus

NADPH oxidase might negatively feedback regulate the generat...

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中国植物病理学会2015年学术年会

作者： MEI Peng-ying ZHOU Chang-yong citrus Research Institute Southwest Universtity

citrus canker,caused by Xanthomonas citri ***(Xcc),is a devastating bacterial disease threatening citrus industry ***-adenine dinucleotide phosphate oxidase(NADPH oxidase),encoded by rboh(respiratory burst oxidase homologues),transfers electrons from NADPH to molecular oxygen to generate O ?-,which is the first step in the formation of HOin plant-pathogen interaction *** studies have shown that the concentrations of HOvary a lot in various citrus varieties when challenged with Xcc,and the H O level has been correlated with disease *** paper aimed to analyze the average 2(cycle threshold) values of rboh expression in kumquat(Fortunella margarita),Ponkan(citrus reticulata Ponkan),sweet orange(*** Osbeck) and grapefruit(*** Duncan),respectively after infection with Xcc,and to observe the potential relationship between the rboh gene expression in the leaves of these four citrus varieties and the resistance level differences among those *** results showed that the average2 values of rboh expression decreased in all four varieties after inoculation with ***,the rboh in four varieties expressed within 1 dpi(days post inoculation) and the concentrations of HO were monitored,while within 8 12 dpi,the rboh expression decreased and the HO concentration surged a second time in kumquat,Ponkan and sweet ***,in grapefruit,the rboh expression increased but the HO concentration decreased within 8 12 ***,the average 2values of rboh expression,both before and after inoculation,were listed from high to low;grapefruit,sweet orange,Ponkan and kumquat,contrary to that in H O ***,it was suggested that the rboh gene might be a negative feedback regulator of the HO generation.

关键词： NADPH oxidase rboh citrus canker hydrogen peroxide

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